Invest Clin 63(4): 353 - 362, 2022 https://doi.org/10.54817/IC.v63n4a03

Corresponding author. Wangjun Gao. Department of General Surgery, Ankang Central Hospital, Ankang, China.

E-mail: ziyaan767193@163.com

The effects of curcumin on the biological

behavior of colorectal cancer cells through

the JAK/STAT3 and RAS/MAPK/NF-κB

pathways.

Zhe Yang

, Rui Zhao

and Wangjun Gao

Department of Radiotherapy, Baoji Municipal Central Hospital, Baoji, China.

Department of Anorectal Surgery, Baoji Municipal Central Hospital, Baoji, China.

Department of General Surgery, Ankang Central Hospital, Ankang, China.

Keywords: human colorectal cancer cells; HCT116 cells; growth cycle; proliferation;

apoptosis.

Abstract. The purpose of this work was to investigate the effects of curcumin

on the biological behavior of colorectal cancer cells through the JAK/STAT3 and

RAS/MAPK/NF-κB pathways. Human colorectal cancer HCT116 cells were cultured

and divided into a control group and low, medium and high-dose curcumin groups

(n =5). HCT116 colorectal cancer cells became long-growing cells after incubation

and culture at 37°C. The control group was treated with 15μL phosphate-buffered

saline, and the low-dose, medium-dose and high-dose curcumin groups were treated

with 20, 40 and 80μmol/L curcumin, respectively. All groups were treated with rel-

evant drug intervention, digested and centrifuged for 48h, washed twice with a PBS

solution, centrifuged at 1000 rpm for 3 min, and the cells precipitated. The prolif-

eration, apoptosis and growth cycle of cells in each group were observed, and the ex-

pressions of the JAK/STAT3 and RAS/MAPK/NF-κB pathways and related proteins in

each group were studied. Compared with the curcumin low-dose and medium-dose

groups, the proliferation ability of the curcumin high-dose group was significantly

decreased (P<0.05). When the low-dose and medium-dose curcumin groups were

compared with the high-dose curcumin group, the apoptosis ability was significantly

increased (P<0.05). When the low-dose and medium-dose curcumin groups were

compared, the growth ratio of the G0/G1 phase in the high-dose curcumin group

was significantly increased, and the percentage of the S phase was significantly de-

creased (P<0.05). Compared with the curcumin low-dose and medium-dose groups,

the expression of JAK-STAT3 and RAS/MAPK/NF-κB pathway in the curcumin high-

dose group was significantly decreased (P<0.05). The protein expressions of STAT3,

RAS, P-P38 and P65 in the curcumin high-dose group were significantly lower than

those in the curcumin low-dose and medium-dose groups (P<0.05). Curcumin can

inhibit the expression of JAK/STAT3 and RAS/MAPK/NF-κB pathways, block the

growth cycle, and inhibit the proliferation and induce apoptosis of colorectal cancer

cells, providing a new idea for the clinical treatment of colorectal cancer.

354 Yang et al.

Investigación Clínica 63(4): 2022

Los efectos de la curcumina en el comportamiento biológico de

las células del cáncer colorrectal mediante las vías JAK/STAT3

y RAS/MAPK/NF-KB.

Invest Clin 2022; 63 (4): 353 – 362

Palabras clave: células de cáncer colorrectal humano; células HCT116; ciclo de

crecimiento; proliferación; apoptosis.

Resumen. El objetivo del presente trabajo fue investigar los efectos de la

curcumina en el comportamiento biológico de las células del cáncer colorrec-

tal mediante el estudio de las vías JAK/STAT3 y RAS/MAPK/NF-KB. Las células

del cáncer colorrectal humano HCT116 se cultivaron y dividieron en un grupo

control y en grupos con dosis baja, media y alta (n = 5) de curcumina. Las cé-

lulas de cáncer colorrectal HCT116 se convirtieron en células de crecimiento

prolongado después de la incubación y cultivo a 37°C. El grupo de control se

trató con 15 μL de solución tampón fosfato salina (PBS) y los grupos de curcu-

mina de dosis baja, media y alta se trataron con 20, 40 y 80 μmol/L de curcu-

mina, respectivamente. Todos los grupos fueron tratados con la intervención

farmacológica pertinente, digeridos y centrifugados durante 48 horas, lavados

dos veces con solución de PBS, centrifugados a 1000 rpm durante 3 minutos,

y las células precipitadas. Se observaron la proliferación, la apoptosis y el ciclo

de crecimiento de las células de cada grupo, y fueron estudiados las expresio-

nes de las vías JAK/STAT3, RAS/MAPK/NF-KB y proteínas relacionadas en cada

grupo. Comparado con los grupos de la dosis baja y media de la curcumina,

disminuyó obviamente la capacidad de proliferación del grupo de la dosis alta

de la curcumina (P<0,05). Comparado con los grupos de la dosis baja y media

de la curcumina, aumentó de modo significativo la capacidad de la apoptosis

del grupo de la dosis alta de la curcumina (P<0,05). Comparado con los grupos

de la curcumina de dosis baja y media, aumentó obviamente la proporción del

crecimiento de la fase G0/G1 en el grupo de la curcumina de dosis alta y el por-

centaje de la fase S disminuyó considerablemente (P<0,05). Las expresiones

proteicas STAT3, RAS, P-P38 y P65 en el grupo de la dosis alta de la curcumina

fueron evidentemente más bajas que las de los grupos de la dosis baja y media

de la curcumina (P<0.05). La curcumina puede inhibir la expresión de las vías

JAK/STAT3 y RAS/MAPK/NF-KB, bloquear el ciclo del crecimiento y luego in-

hibir la proliferación e inducir apoptosis de las células del cáncer colorrectal,

lo que brinda una nueva idea para el tratamiento clínico del cáncer colorrectal.

Received: 30-04-2022 Accepted: 04-06-2022

INTRODUCTION

Colorectal cancer is a common gastroin-

testinal malignancy in clinical practice, ran-

king third among malignancies worldwide,

and its mortality rate is second only to liver

cancer and lung cancer

. Data survey shows

that the incidence and mortality of colorectal

cancer are on the rise, and its mortality rate

ranks second among malignant tumors in de-

veloped countries, seriously threatening hu-

man health and quality of life

. Studies have

The effects of curcumin on the biological behavior of colorectal cancer cells 355

Vol. 63(4): 353 - 362, 2022

found that the incidence of colorectal cancer

is gradually rising in younger people, and the

prognosis of patients underyears old-old is

poor

. Surgical resection is currently the pri-

mary method treatingt for colorectal cancer

in clinical practice. Patients in advanced sta-

ges are mostly treated with radiotherapy and

chemotherapy, but the treatment effect of co-

lorectal cancer is not ideal; and the progno-

sis of the patients is poor, and most patients

suffer from recurrence and metastasis

. Cur-

cumin is a natural and effective chemical

component in plants. It mainly exists in the

roots and stems of turm,eric and it is insolu-

ble in water. It can change with the change of

acid and alkali conditions. Curcumin is widely

used in food production as a preservative, co-

lorant, etc., and also plan essentialtant role

in anti-tumor, hypolipidemic, anti-oxidation

and anti-arteriosclerosis

. Jabus kinases/sig-

nal and activator of the transcription Jak-stat

pathway is involved in the cycle, cell transfor-

mation and apoptosis of tumor cells

. It was

found

that Rat sarcoma (RAS) protein can

activate the mitogen activated protein kina-

se (MAPK) and the nuclear factor (NF-κB)

pathway. It can induce the proliferation of he-

patocellular carcinoma cells and strengthen

their invasion ability. However, curcumin has

been rarely studied in the JAK/STAT3 and

RAS/MAPK/NF-κB pathways of colorectal

cancer. Therefore, this study aimed to explore

the effects of curcumin on the biological be-

havior of colorectal cancer cells through the

JAK/STAT3 and RAS/MAPK/NF-κB pathways.

MATERIAL AND METHODS

Experimental materials

Human colorectal cancer HCT116 cells

were purchased from Hubei Punosai Life

Science and Technology Co., LTD., and were

uniformly cryopreserved by the experimen-

tal center of our hospital.

Experimental instruments and reagents

Curcumin (Beijing Green Heng Xing

Biotechnology Co., LTD.), PBS buffer (Bei-

jing Standard Technology Effective Com-

pany), medium (Shanghai Biotechnology

Co., LTD.), fetal calf serum (Jiangsu Ke Wei

Biotechnology Co., LTD.), dimethyl sulfoxi-

de (Chengdu Medical Technology Co., LTD.),

automatic labeling instrument (Shanghai

Molecular Instrument Co., LTD.,), protein

extraction kit (Beijing Solaibao Technology

Co., LTD.), flow cytometer (Shanghai Huan-

lian Medical Device Co., Ltd.).

Cell Grouping

Colorectal cancer HCT116 cells were

placed in 10% fetal bovine serum and cul-

tured in a cell incubator with 5% CO

and

37°C. When the cells were fused to 90%, a

trypsin solution was given for digestion and

passage treatment. When the passage rea-

ched the third generation, cells with long-

term growth were selected for experiment.

In the process of culture, the growth state

of cells was observed, and the conventional

fluid was changed according to its state. Co-

lorectal cancer HCT116 cells were divided

into control group, and a curcumin low-

dose, medium-dose and high-dose groups.

The control group was treated with 15μL

phosphate buffered saline, and the low-do-

se, medium-dose and high-dose curcumin

groups were treated with 20, 40 and 80

μmol/L curcumin, respectively, to observe

the proliferation and apoptosis of cells in

each group. The expression of JAK/STAT3,

RAS/MAPK/NF-κB pathway and related pro-

teins were studied in each group.

EXPERIMENTAL METHODS

Cell proliferation experiment

The cell cycle is the most important

entity for cell survival. Abundant factors

and proteins in positive or negative maps at

multiple points and bottlenecks, precisely

and harmoniously regulate and control this

cycle. In fact, there are a variety of genes in

cells that encode proteins needed to control

the cell cycle. Although the cell cycle is mo-

nitored and inspected at several stations,

356 Yang et al.

Investigación Clínica 63(4): 2022

this adjustment takes place especially at two

points with extraordinary intensity and care.

The cell, first, decides to replicate its own

DNA, and second, to initiate mitotic divi-

sion. These steps are in the realm of passing

from G1 to S and from G1 to M.

The cells in each group were successi-

vely inoculated into a 96-well culture plate,

and the cell density was adjusted to 10×10

mL. The cells were observed for 5h, and the

corresponding intervention was given to

each group. Cultured again for 36 h, the

MTT assay was used to observe the situation

of each group of cells, and the proliferation

of each group was plotted. The experiment

was repeated 3 times.

Flow cytometry detection

Colorectal cancer HCT116 cells with

a concentration of 2×10

/L were inocula-

ted into 6-well culture plates for 12 h. The

control group was given routine culture, and

the curcumin low-dose, medium-dose and

high-dose groups were given 20, 40 and 80

μmol/L curcumin for intervention. After 48

h of intervention, the cells were collected

and centrifuged at 1000 rpm for 10 min.

The cells were washed with PBS twice, and

1ml of precooled 70% ethanol was used for

beating and dispersing. The cells were fixed

overnight at -20°C, and then mixed with 0.8

μg/mL. The cells were incubated at room

temperature for 30 min without light.

Western blot assay

Western blot analysis was performed

essentially according to standard protocol.

Briefly, the cells were solubilized in lysis

buffer (50 mM Tris, 100 mM NaCl, 2.5 mM

EDTA, 1% Triton X-100, 1% Nonidet P-40, 2.5

mM Na3O4V, 25 μg/mL aprotinin, 25 μg/mL

leupeptin, 25 μg/mL pepstatin A, and 1 mM

phenylmethylsulfonyl fluoride). After clarifi-

cation at 10,000g for 15 minutes, the super-

natant was used for Western blot analysis. In

all analyses, protein concentration, determi-

ned by the Bio-Rad Protein Assay kit (Bio-

Rad, Hercules, CA), was standardized among

the samples. Aliquots of cell lysates con-

taining 50 μg of protein were separated by

sodium dodecyl sulfate–polyacrylamide gel

electrophoresis. After electrophoresis, pro-

teins were transferred electrophoretically

onto supported nitrocellulose membranes

(Osmonics, Gloucester, MA). Membranes

were incubated for 1 hour at room tempe-

rature with blocking buffer, TBS-T (20 mM

Tris, pH7.6, 100 μMNaCl, 0.1% Tween-20)

and 5% nonfat dry milk with gentle agita-

tion. After washing the membranes with

TBS-T, they were incubated overnight at 4°C

in TBS-T buffer containing antibody dilution

buffer as suggested by the manufacturer and

with antibodies (1:1000 dilution) to CD44,

CD166 (Santa Cruz Biotechnology, Santa

Cruz, CA), or epidermal growth factor re-

ceptor (EGFR; Cell Signaling, Beverly, MA).

The membranes were washed three times

with TBS-T and subsequently incubated with

appropriate secondary antibodies (1:5000

dilutions) in TBS-T containing 5% milk for

1 to 2 hours at room temperature with gent-

le agitation. The membranes were washed

again with TBS-T, and the protein bands were

visualized by enhanced chemiluminescence

(ECL) detection system (Amersham, Pisca-

taway, NJ). The membranes containing the

electrophoresed proteins were exposed to

X-Omat film (Sigma-Aldrich, St Louis, MO).

The membranes were stripped (twice × for

15 minutes at 55°C) in stripping buffer con-

taining 100 mM 2-mercaptoethanol, 2% so-

dium dodecyl sulfate, and 62.5 mM Tris-HCl

pH 6.7, and reprobed for β-actin using the

corresponding antibodies, which were used

as a loading control. All Western blots were

performed at least three times for each expe-

riment

. The method used to identify prote-

ins in the membrane is the blotting method.

In this technique, protein bands are transfe-

rred from the gel to a nitrocellulose mem-

brane that can bind and stabilize proteins.

To do this, by blotting, the protein molecules

were removed from the gel and placed in the

same position on the surface of the membra-

ne, so we could easily study them, separate

The effects of curcumin on the biological behavior of colorectal cancer cells 357

Vol. 63(4): 353 - 362, 2022

them, and finally use them. Specific ligands

or substrates were used to detect proteins or

enzymes transferred to the membrane. Anti-

bodies were also used to specifically detect

proteins in the membrane.

All groups were treated with relevant

drug intervention, digested and centrifuged

for 48h, washed twice with PBS solution,

centrifuged at 1000 rpm for 3min, and the

cells precipitated. The protein was extracted,

200μL lysate was put into the cell sample,

let stand and mix. The BCA method was used

to determine protein content according to

the kit instructions. After gel preparation,

sample loading, electrophoresis, membrane

transfer, elution, sealing, primary antibody

incubation, membrane washing, secondary

antibody incubation and membrane washing

again, the protein expression was calculated

by the Invitrogen

/Thermo Fischer Scienti-

fic, USA, technique.

Apoptosis methodology

There are two pathways for cell apopto-

sis, intrinsic and external. For this study, the

TUNEL method was used to evaluate apop-

tosis

To diagnose apoptosis, the TUNEL kit

(insitu cell death detection kit, POD, Roche

company, made in Germany) (Cat. No. 11

684 817 910) was used. The TUNEL techni-

que was performed as follows:

1. First, the prepared sections were

washed with para-proteinase K after paraffi-

nization and irrigation, and after incubation,

they were washed with buffer phosphate so-

lution for 30 minutes at 37°C.

2. Tissue sections were stained with 50

μl of TUNEL reaction solution for 37 minutes

at 37°C, then washed with buffer phosphate

solution.

3. At this stage, the sections were then

washed with a converter solution (50 μL)

for 30 minutes at 37°C with buffer phospha-

te solution and then mixed with a diamino

benzidine solution for 20 minutes and then

stirred for 25 minutes. They were incubated

again at 37°C.

4. The sections were then washed three

times with phosphate solution and batoloi-

din blue, for two minutes each time.

Cells were manually examined in 10

random high-power (×100 magnification)

fields (>1000 cells) and the apoptosis index

(AI) was determined as follows: AI = number

of positively stained cells/total number of

cells counted.

Statistical methods

The SPSS20.0 software package was

used for statistical analysis of the study data,

and the measurement data was expressed as

means ± standard deviation (

sx ±

). One

way-ANOVA was used for comparison bet-

ween groups. P<0.05 was taken as the sta-

tistical standard.

RESULTS

Comparison of proliferation ability

of different groups of cells

Compared with the control group, the

proliferation ability of the medication group

was significantly decreased, with statistical

significance (P<0.05); compared with the

low-dose and medium-dose curcumin groups,

the proliferation ability of the high-dose cur-

cumin group was significantly decreased,

with statistical significance (P<0.05), as

shown in Table 1.

Comparison of apoptosis in different

groups

Compared with the control group, the

apoptotic ability of the medication group

was significantly increased (P<0.05); com-

pared with the low-dose and medium-dose

curcumin groups, the apoptotic ability of the

high-dose curcumin group was significantly

increased (P<0.05), as shown in Table 2.

Comparison of cell growth cycles

of different groups

Compared with the control group, the-

re was no significant difference in G2/M

phase (P>0.05), but the growth ratio of G0/

358 Yang et al.

Investigación Clínica 63(4): 2022

G1 phase in the treatment group was signi-

ficantly increased, and the percentage of S

phase was significantly decreased, the diffe-

rence was statistically significant (P<0.05).

Compared with the low-dose and medium-do-

se curcumin groups, the growth ratio of G0/

G1 phase in the high-dose curcumin group

was significantly increased. The percentage

of S stage decreased significantly, with sta-

tistically significant difference (P<0.05), as

shown in Table 3.

Comparison of JAK-STAT3 and RAS/

MAPK/NF-κB pathway expression in

different groups

Compared with the control group, the

expression of JAK-STAT3 and RAS/MAPK/

NF-κB pathway in the treatment group was

significantly decreased, and the differences

were statistically significant (P<0.05).The

expression of JAK-STAT3 and RAS/MAPK/

NF-κB pathway (μg) was significantly decrea-

sed in the high-dose curcumin group, and

the differences were statistically significant

(P<0.05), as shown in Table 4.

Comparison of related protein expression

in different groups

Compared with the control group, the

protein expressions (μg) of STAT3, RAS, P-P38

and P65 in the medication group were signi-

ficantly decreased, with statistical significance

(P<0.05). Compared with the low-dose and

medium-dose curcumin groups, the protein ex-

pressions of STAT3, RAS, P-p38 and P65 in the

high-dose curcumin group were significantly

decreased. The difference was statistically sig-

nificant (P<0.05), as shown in Table 5.

Table 1

Comparison of migration ability (proliferation) of different groups of cells (

sx ±

)

Group Number of stems

proliferation ability（%）

control group 5 96.44 ± 3.15

low-dose curcumin group 5 76.16 ± 5.33

medium-dose curcumin group 5 59.13 ± 4.16

high-dose curcumin group 5

45.11 ± 3.85

*#△

F 139.29

＜0.001

Compared with control group, *P<0.05; compared with curcumin low-dose group,

P<0.05; compared with cur-

cumin medium-dose group,

△

P<0.05. P-Value based on One Way- ANOVA. Number of stems: number of groups

of cells.

Table 2

Comparison of apoptosis in different groups (

sx ±

)

Group Number of stems Apoptosis (%)

control group 5 5.23 ± 1.32

low-dose curcumin group 5 14.35 ± 1.22*

medium-dose curcumin group 5 25.76 ± 1.68*

high-dose curcumin group 5

35.79 ± 2.11*

#△

F 337.77

P <0.001

Compared with control group, *p < 0.05; compared with curcumin low-dose group,

p< 0.05; compared with curcumin

medium-dose group,

△

p< 0.05. P-Value based on One Way- ANOVA. Number of stems: number of groups of cells.

The effects of curcumin on the biological behavior of colorectal cancer cells 359

Vol. 63(4): 353 - 362, 2022

Table 3

Comparison of cell growth cycles of different groups (

sx ±

)

Group

Number

of stems

G0/G1(%) S(%) G2/M(%)

control group 5 33.26 ± 2.19 53.76 ± 3.11 11.52 ± 3.16

low-dose curcumin group 5 45.38 ± 2.56* 41.35 ± 2.94* 11.31 ± 2.25

medium-dose curcumin group 5 59.67 ± 3.12*

30.85 ± 3.16*

10.23 ± 1.35

high-dose curcumin group 5

71.31 ± 3.15*

#△

20.33 ± 2.08*

#△

9.22 ± 1.64

F 177.61 125.62 1.00

P <0.001 <0.001 0.418

Compared with control group, *p<0.05; compared with curcumin low-dose group,

p<0.05; compared with cur-

cumin medium-dose group,

△

p<0.05. P-Value based on One Way- ANOVA. Number of stems: number of groups of

cells.

Table 4

Comparison of JAK-STAT3 and RAS/MAPK/NF-κB pathway expression (μg) in different groups (

sx ±

)

Group

Number

of stems

JAK-STAT3 RAS p38MAPK NF-κB

control group 5 1.06±0.09 1.15±0.26 0.98±0.02 0.85±0.16

low-dose curcumin group 5 0.71±0.11* 0.73±0.09* 0.75±0.11* 0.54±0.05*

medium-dose curcumin group 5 0.51±0.06*

0.53±0.05*

0.42±0.07*

0.40±0.05*

high-dose curcumin group 5

0.31±0.02*

#△

0.29±0.04*

#△

0.26±0.03*

#△

0.22±0.04*

#△

F 32.07 33.24 114.71 43.99

P <0.001 <0.001 <0.001 <0.001

Compared with control group, *p<0.05; compared with curcumin low-dose group,

p<0.05; Compared with cur-

cumin medium-dose group,

△

p<0.05. P-Value based on One Way- ANOVA. Number of stems: number of groups of

cells. Unit of protein values: μg.

Table 5

Comparison of related protein expression (μg) in different groups (

sx ±

)

Group

Number

of stems

STAT3 ras p-p38 p65

control group 5 1.31 ± 0.35 1.53 ± 0.41 1.35 ± 0.31 1.45 ± 0.15

low-dose curcumin

group

0.92 ± 0.19*

1.02 ± 0.19*

0.85 ± 0.08*

0.73 ± 0.11*

medium-dose

curcumin group

0.59 ± 0.06*

0.62 ± 0.08*

0.51 ± 0.06*

0.52 ± 0.05*

high-dose curcumin

group

0.29 ± 0.02*

#△

0.28 ± 0.01*

#△

0.26 ± 0.01*

#△

0.21 ± 0.03*

#△

F 23.64 27.48 41.90 146.12

P <0.001 <0.001 <0.001 <0.001

Compared with control group, *p < 0.05; compared with curcumin low-dose group,

p< 0.05; compared with cur-

cumin medium-dose group,

△

p< 0.05. P-Value based on One Way- ANOVA. Number of stems: number of groups of

cells. Unit of protein values: μg.

360 Yang et al.

Investigación Clínica 63(4): 2022

DISCUSSION

With the changes of diet structure and

the improvement of living standards, the in-

cidence of colorectal cancer has been high,

and is closely related to genetics, the envi-

ronment and diet. Data survey shows that

the incidence of colorectal cancer in big

cities in China is higher than that in other

cities, and it shows an increasing trend year

by year. Colorectal cancer has become the

most common and fastest growing malig-

nant tumor in China

. Surgery combined

with radiotherapy and chemotherapy is com-

monly used in the clinical treatment of co-

lorectal cancer, but about 50% of patients

will have recurrence and metastasis after

surgery, which will eventually lead to death

of patients

. The application of chemothe-

rapy drugs can lead to serious toxic and side

effects in patients, waste medical resources,

and bring serious economic burden to fami-

lies and society. Therefore, finding effective

and safe drugs to treat colorectal cancer has

become the focus of clinical research.

Traditional Chinese medicine treatment

has been gradually applied in clinical practi-

ce, and has unique advantages, and has be-

come an important part of clinical tumor

treatment. Studies have found

that curcu-

min widely exists in turmeric, which can be

combined with chemoradiotherapy drugs to

effectively reduce the toxic and side effects

of chemotherapy drugs, the dosage of the-

se drugs, and to improve the prognosis and

the quality of life of patients. Curcumin has

been applied to thyroid cancer cells, and in-

hibition of thyroid cancer cell proliferation

was observed, which may be related to the

inhibition of p-MTOR and P-S6K proteins in

thyroid cancer

. Another study in thyroid

cancer found that curcumin could down-re-

gulate the expressions of cy-Clinbl and Bcl-

XL, and then inhibit thyroid cancer cells,

thereby promoting their apoptosis

. The re-

sults were similar to those of this group. The

cell growth cycle is the most important step

in tumorigenesis and development. Foreign

studies have found that when curcumin is

applied to colorectal cancer cells, it is found

that G1 block occurs, and then the apopto-

sis specific DNA is delayed, suggesting that

curcumin can regulate the growth cycle of

colorectal cancer cells

. In this experiment,

compared with curcumin low-dose and me-

dium-dose groups, the proliferation ability of

curcumin high-dose group was significantly

decreased, and the apoptosis ability was sig-

nificantly increased (P<0.05). These results

suggest that curcumin can inhibit the proli-

feration of colorectal cancer cells, block the

growth of colorectal cell cycle, and induce

their apoptosis.

During the occurrence and development

of gastric cancer, the Jak-Stat3 pathway and

the RAS-MAPK pathway have been successi-

vely activated and have complex interactions,

which are closely related to gastric cancer.

MAPK is a core member of the RAS-MAPK

pathway, which can transmit extracellular

signals to the nucleus by activating MAPK,

leading to activation of phosphoamino acid

residues of Jun, FOS and other transcrip-

tion factors in the nucleus, thus regulating

gene expression and finally leading to cell

growth and differentiation

. The terminal

of STAT3-C has a serine residue, and Ser727

is the phosphorylation site of MAPK, indica-

ting that STAT3 is closely related to p38MA-

PK. Studies have confirmed that STAT3 has

a certain correlation with p38MAPK in gas-

tric cancer, and the expression of STAT3 can

affect the expression of p38MAPK, which in-

creases with the increase of STAT3

. RAS

exists in the form of binding proteins in DNA

and mutates in tumors, thus affecting the

protein activity of RAS and accelerating the

proliferation, migration and invasion of tu-

mors, and has become an important target

of cancer

. It was found that MAPK/NF-κB

can act as a downstream pathway of RAS, lea-

ding to RAS activation, which in turn activa-

tes MAPK enzyme, resulting in transfer and

phosphorylation of Raf molecules downs-

tream of RAS, thereby activating MAPK, and

activating NF-κB in the nucleus, amplifying

The effects of curcumin on the biological behavior of colorectal cancer cells 361

Vol. 63(4): 353 - 362, 2022

RAS activity. Ultimately, the inflammatory

response is accelerated

. In this study, the

expressions of JAK-STAT3 and RAS/MAPK/

NF-κB pathways were significantly decreased

in the curcumin high-dose group, and the

protein expressions of STAT3, RAS, P-P38

and P65 were significantly decreased in cur-

cumin low-dose and medium-dose groups (P

< 0.05). These results suggest that curcu-

min can inhibit the expression of JAK-STAT3

and RAS/MAPK/NF-κB pathways, further in-

hibit related proteins, and prevent further

proliferation of colorectal cancer cells.

In this experimental study, curcumin

could inhibit the proliferation and induce

apoptosis of colorectal cancer cells by in-

hibiting the expression of JAK/STAT3 and

RAS/MAPK/NF-κB pathways and block the

growth cycle, providing a new idea for clini-

cal treatment of colorectal cancer.

Conflict of interest

The manuscript has no conflict of in-

terest.

Author’s ORCID numbers

• Zhe Yang: 0000-0002-3176-5853

• Rui Zhao: 0000-0001-7973-2762

• Wangjun Gao: 0000-0003-2560-441X

Limitation

Low number of groups of cells.

Authors Contribution

ZY and RZhao collected the samples.

ZY and WG analysed the data. RZ and WG

conducted the experiments and analyzed the

results. All authors discussed the results and

wrote the manuscript.

Funding

None.

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